This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hanawa, H. Articles by Nienhuis, A. W. Search for Related Content PubMed PubMed Citation Articles by Hanawa, H. Articles by Nienhuis, A. W.

 Previous Article  |  Next Article 

Journal of Virology, July 2005, p. 8410-8421, Vol. 79, No. 13
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.13.8410-8421.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Mobilization and Mechanism of Transcription of Integrated Self-Inactivating Lentiviral Vectors

Hideki Hanawa, Derek A. Persons, and Arthur W. Nienhuis^*

Division of Experimental Hematology, Department of Hematology/Oncology, St. Jude Children's Research Hospital, 332 N. Lauderdale, Memphis, Tennessee 38105

Received 13 December 2004/ Accepted 11 March 2005

Permanent genetic modification of replicating primitive hematopoietic cells by an integrated vector has many potential therapeutic applications. Both oncoretroviral and lentiviral vectors have a predilection for integration into transcriptionally active genes, creating the potential for promoter activation or gene disruption. The use of self-inactivating (SIN) vectors in which a deletion of the enhancer and promoter sequences from the 3' long terminal repeat (LTR) is copied over into the 5' LTR during vector integration is designed to improve safety by reducing the risk of mobilization of the vector genome and the influence of the LTR on nearby cellular promoters. Our results indicate that SIN vectors are mobilized in cells expressing lentiviral proteins, with the frequency of mobilization influenced by features of the vector design. The mechanism of transcription of integrated vector genomes was evaluated using a promoter trap design with a vector encoding tat but lacking an upstream promoter in a cell line in which drug resistance depended on tat expression. In six clones studied, all transcripts originated from cryptic promoters either upstream or within the vector genome. We estimate that approximately 1 in 3,000 integrated vector genomes is transcribed, leading to the inference that activation of cryptic promoters must depend on local features of chromatin structure and the constellation of nearby regulatory elements as well as the nature of the regulatory elements within the vector.

---

* Corresponding author. Mailing address: St. Jude Children's Research Hospital, 332 N. Lauderdale, Mail Stop #272, Memphis, TN 38105. Phone: (901) 495-3301. Fax: (901) 525-2720. E-mail: arthur.nienhuis{at}stjude.org.

---

Journal of Virology, July 2005, p. 8410-8421, Vol. 79, No. 13
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.13.8410-8421.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Uchida, N., Washington, K. N., Hayakawa, J., Hsieh, M. M., Bonifacino, A. C., Krouse, A. E., Metzger, M. E., Donahue, R. E., Tisdale, J. F. (2009). Development of a Human Immunodeficiency Virus Type 1-Based Lentiviral Vector That Allows Efficient Transduction of both Human and Rhesus Blood Cells. J. Virol. 83: 9854-9862 [Abstract] [Full Text]  
• Zong, Y., Zhou, S., Fatima, S., Sorrentino, B. P. (2006). Expression of Mouse Abcg2 mRNA during Hematopoiesis Is Regulated by Alternative Use of Multiple Leader Exons and Promoters. J. Biol. Chem. 281: 29625-29632 [Abstract] [Full Text]